柯里拉京对人喉癌Hep-2细胞生长的研究

Study on apoptosis of laryngeal carcinoma Hep-2 Induced by corilagiln

目的:研究柯里拉京(Corilagiln)对人喉癌细胞Hep-2的抑制生长作用及凋亡诱导作用。方法 :四亚基噻唑蓝(MTT)法测定柯里拉京对细胞生长的抑制作用,流式细胞仪技术(FCM)观察细胞周期及凋亡;免疫印记法(Western-blot)测定Bax及bcl-2表达,荧光定量PCR法测量人喉癌Hep-2细胞增殖诱导配体(aproliferation-inducing ligand,APRIL)的表达量。结果:2.5～20μg/ml的柯里拉京均能抑制人喉癌细胞Hep-2生长,并有明显的时间和剂量依赖性,2.0μg/ml的柯里拉京作用72h后对人喉癌细胞Hep-2生长抑制作用达到73%;流式细胞仪检测不同浓度柯里拉京作用72h后的细胞凋亡率分别为13.80%±2.32%,21.72%±1.51%,37.43%±1.06%和54.91%±2.40%;经柯里拉京作用后,免疫印记法显示柯里拉京上调Bax,同时降低bcl-2表达,使Bax/bcl-2比值增加;柯里拉京作用人喉癌Hep-2细胞后其增殖诱导配体的mRNA表达量随时间的延长和浓度的增加而升高,48 h后各浓度与空白组比较差异有统计学意义(P<0.05)。结论:柯里拉京在体外能明显抑制人喉癌细胞Hep-2生长,抑制其生长的机制可能是诱导细胞凋亡。

To determine the effect of corilagiIn on the growth in human Laryngeal carcinoma Hep-2 ceils. Methods ： Using MTr as- say, the growth inhibiting rate of cerilagiln on Hep-2 cells was dedected. Fluorescehce microscope and flow cytometry（FCM） were applied to demonstrate the presence of apoptosis and cell cycleo The expression of Bax and bcl-2 was analyzed with Western-blot. the mRNA expression of a proliferation-inducing ligand （APRIL） APRIL in the Hep-2 ceils was detected by real-time fluorescent quantitative PCR. Resulls： Cori- lagiln inhibited the growth of Hep-2 ceils in a time- and dose-dependent manner. The apoptotic rates at 2.5.5.0. 10.0 and 20.0 μg/ml for 72h were 13.81%± 2.32% ,21.82% ＋ 1.51% ,37.53% ± 1.06% and 54. 90% ± 2. 40%, respectively, which were all significantly higher than those of the control group （ P 〈 0. 05 ）. Western-blot showed that Bax expression was up-regulated while bcl-2 expression was down-regulated in the Hep-2 ceils treated with Corilagiln, thus increasing the ratio of Bax/bcl-2. The mRNA level of a prollferation-inducing ligand APRIL in the Hep-2 cells raised with time prolonging and concentration increasing, and was significantly higher than the blank group after 48 h （ P 〈 0.05）. Conclusion： Cofilagiln can inhibit the proliferation of human Laryngeal carcinoma Hep-2 cells in a time- and dase-de- pendent manner and the mechanism of growth inhibition of human Laryngeal carcinoma ceils may be related to the induction of apoptosis.